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目的 :研究类胰蛋白酶抑制剂 (TPI)对人大肠肥大细胞释放组胺的影响。方法 :大肠组织经胶原酶和透明质酸酶消化后 ,将细胞成分用全HBSS重新悬浮。肥大细胞在LP4试管中于37℃条件下与各种刺激剂反应 15min而完成激发过程。激发液中的组胺水平用以玻璃纤维为基础的荧光方法测定。结果 :4种TPI中 ,高浓度的亮抑酶肽素和鱼精蛋白可刺激人大肠肥大细胞释放组胺 ;而TLCK和乳铁蛋白则无明显的刺激作用。 4种TPI均可以剂量依赖的方式抑制抗IgE抗体诱导的大肠肥大细胞释放组胺。最大浓度的亮抑酶肽素(2 0 0mmol/L)、TLCK(10 0mmol/L)、乳铁蛋白 (30mmol/L)和鱼精蛋白 (10 0mg/L) ,可分别抑制 4 8.7%、36 .7%、4 0 .2 %和 34.1%的组胺释放。在 37℃条件下 ,将 4种TPI同大肠肥大细胞预培养 2 0min ,与未进行预培养相比较 ,它们对抗IgE抗体诱导的组胺释放无明显改变。 4种TPI还可抑制CI诱导的组胺释放 ,抑制范围在 2 5 %～ 32 %之间。与抗IgE抗体诱导的组胺释放则不同 ,与大肠肥大细胞预培养 2 0min ,与未进行预培养相比较 ,亮抑酶肽素和TLCK对CI诱导的组胺释放的抑制作用明显增强 ;而鱼精蛋白则无此作用。结论 :我们首次发现TPI可抑制人大肠肥大细胞IgE抗体依赖和非IgE抗体依赖的组胺释放 ,提示TPI可望成为炎症性肠
Objective: To study the effect of tryptase inhibitor (TPI) on histamine release from human large intestine mast cells. METHODS: After digestion by collagenase and hyaluronidase in colorectal tissues, the cell fraction was resuspended with whole HBSS. Mast cells in the LP4 tube at 37 ℃ conditions with a variety of stimuli for 15min to complete the excitation process. Histamine levels in the excitation fluid were determined using a glass fiber-based fluorescence method. RESULTS: High concentration of aprotinin and protamine stimulated the release of histamine in human large intestine mast cells in four TPIs, whereas TLCK and lactoferrin had no obvious stimulating effects. All four TPIs inhibited histamine release from large intestine mast cells induced by anti-IgE antibodies in a dose-dependent manner. The highest concentrations of bright aprotinin (200 mmol / L), TLCK (10 0 mmol / L), lactoferrin (30 mmol / L) and protamine (10 0 mg / L) 36.7%, 40.2% and 34.1% histamine release. Four kinds of TPIs were pre-incubated with large intestine mast cells for 20 min at 37 ℃. Compared with the non-pre-cultured ones, they showed no significant changes in histamine release induced by anti-IgE antibodies. Four TPIs also inhibited the CI-induced histamine release, with inhibition ranging from 25% to 32%. Compared with anti-IgE antibody-induced histamine release, colorectal mast cells were preincubated for 20 min. The inhibitory effect of aprotinin and TLCK on CI-induced histamine release was significantly enhanced compared with that of pre-culture without pre-culture Protamine does not have this effect. CONCLUSIONS: We first found that TPI inhibits IgE antibody-dependent and non-IgE antibody-dependent histamine release in human large intestine mast cells, suggesting that TPI is expected to become an inflammatory bowel